Figure 4. TLR4-KO old mice are protected from adipose tissue inflammation and ER stress. Relative mRNA expressions of ER stress response genes (A) and autophagy genes (B) in adipose tissue of old (20 m) WT and TLR4 deficient mice were analyzed by qRT-PCR. Data represented in bar diagrams are mean + SD value of relative mRNA expression from three independent experiments where total RNA was extracted from the adipose tissue of WT (n=5) and TLR4 deficient (n=5) mice and used as a template for one-step qRT-PCR reaction. (C) Western blot analysis of II/I from SVFs lysates isolated from WT (n=5) and TLR4-KO (n=5) and pooled and treated with either vehicle (DMSO) or Baf (10 nM) for 18h in culture. The density of protein bands from three independent experiments were normalized with α-tubulin and plotted (D). Values were presented as mean + SD of three independent experiments. Significance of difference between means was determined by Student’s t-test and indicated by *p<0.05 and **p<0.01. Symbol # indicated the significance level (p<0.05) of two-way ANOVA analysis for the interaction between treatment (vehicle and Baf) and age factor (WT vs. KO). No significant differences are indicated by ns.